CN105447763A - Cold resistance appraisal method of cotyledon period - Google Patents
Cold resistance appraisal method of cotyledon period Download PDFInfo
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Abstract
The invention relates to the cold resistance appraisal of plants, and particularly provides a cold resistance appraisal method of a cotyledon period. The cold resistance appraisal method of the cotyledon period comprises the following steps: carrying out low-temperature processing on the strong seedlings of the cotyledon periods of a plurality of strains to be tested, wherein the plurality of strains exhibit even growth vigor; then, causing the strains to recover growth under a normal temperature; investigating the states of stems and a situation that cotyledons are damaged; judging a cold damage level according to a standard; calculating the cold resistance index of a variety to be tested; and appraising the cold resistance level according to the standard. The appraisal method disclosed by the invention has the advantages of being simple, visual, reliable and short in periods, can effectively carry out the cold resistance screening of cotton seedling stage germplasm resources and varieties, and paves the way for the further research of the cold resistance breeding and the cold resistance mechanism of the cotton.
Description
Technical field
The present invention relates to the chilling tolerance detaction of plant, being specifically related to a kind of cotton cotyledon phase resists cold authentication method.
Background technology
At occurring in nature, temperature is the important envirment factor affecting plant growth, metabolism.Plant inevitably suffers coercing of various envirment factor, and wherein low temperature is one of main environment stress factors affecting plant life.Low temperature stress makes the viability of crop, the output of crops, situation of growing, photosynthetic rate and mineral element absorption speed etc. be subject to significant suppression, the serious death causing plant.Chilling injury occurs in the warm season of crop growth, and the manifest symptom such as cause crop withered, dead unlike frost and other disaster, chilling temperature may be there is between 15 DEG C and 0 DEG C in damage.The symptom damaged to plants caused by sudden drop in temperature mainly contains: the pathology of surface texture, water logging sample outward appearance, water loss or dehydration, internal discoloration, tissue breakdown, accelerates aging/ethylene production, makes plant metabolites leak and decay faster and shorten the life-span etc.
Cotton originates from the torrid zone, subtropics, belongs to the warm crop of happiness, to cold coerce more responsive, and acclimatization to cold ability.Because spring, autumn air temperature are very unstable, so that cotton in seedling stage, late growth stage usually suffer the harm of low temperature, wherein damage to plants caused by sudden drop in temperature freeze injury seedling stage and occur comparatively frequent.In recent years along with sharply increasing of population and going from bad to worse of weather conditions, the situation that ground striven by grain and cotton is more and more fierce, under the prerequisite ensureing national food security, for ensureing to plant cotton area, the effective way that ground contradiction striven by grain and cotton is alleviated in the transfer of cold and cool area to Inner Mongol, Xinjiang etc. by cotton planting to cultivate cotton cold resistance kind, if the cold resistance of cotton effectively can be improved, prolong growth period, increase the cultivated area of cotton, the per unit area yield of Cotton in China and total product can be increased substantially undoubtedly, contribute to the fiber quality improving cotton simultaneously.
Under cryogenic, identify cotton in seedling stage cold resistance, formulate the appraisement system of a set of cotton in seedling stage cold resistance, theory and practice basis is established in the seed selection for cotton low-temperature tolerant cultivars, for the cold breeding of Resistance Strain of Cotton provides excellent anti-cold germ plasm resource, advance the cold breeding process of Resistance Strain of Cotton.
Summary of the invention
In order to solve the problem, the invention provides a kind of cotton cotyledon phase resists cold authentication method.
The cotton cotyledon phase provided by the invention resists cold authentication method, it is by the healthy and strong seedling sub zero treatment of uniform for growing way many strains strain to be measured cotyledon period, restoration ecosystem at a normal temperature again, then investigate the state of stem and cotyledon to be hurt situation, rank is damaged to plants caused by sudden drop in temperature by the standard determination of table 1, with the formula 1, formula 2 calculates the anti-cold finger number of kind to be measured, differentiates anti-cold rank by the standard of table 2;
The state of described investigation stem and cotyledon are hurt area, specific as follows: the state first investigating stem, be divided into stem rotten disconnected intact with stem, again for the individual plant investigation cotyledon dropping situations that stem is intact, investigate the area that is hurt of the cotyledon that do not come off, it is X that record speck area accounts for this sheet leaf area ratio, and recording total speck area, to account for total leaf area ratio be Y;
Table 1 upland cotton cotyledon period blade damages to plants caused by sudden drop in temperature rank
Chilling injury index (%)=∑ (rank × this grade of strain number)/(8 × total strain number) × 1001
Anti-cold finger number (%)=100-Chilling injury index 2
Table 2 cotyledon period resists cold qualification rank and evaluation criterion
| Anti-cold finger number/% | Anti-cold rank |
| 0.00-29.90 | Cold sensitivity |
| 30.00-49.90 | Not anti-cold |
| 50.00-74.90 | Cold-resistant |
| 75.00-89.90 | Anti-cold |
| ≥90.00 | High resistance is cold |
。
Authentication method of the present invention need have certain sample size, to guarantee the accuracy of qualification result.Wherein, described many strains be 10 strains and more than, preferably 30 strains.
Wherein, described sub zero treatment, for the low temperature that medium cold resistance cotton can be caused to damage to plants caused by sudden drop in temperature, the processing time damages to plants caused by sudden drop in temperature for enough causing, and specifically suitable treatment temperature is 0-15 DEG C, and the processing time is 6-24h; Be preferably 4 DEG C of process 24h.
Wherein, described normal temperature is the temperature that cotton energy normal growth is suitable for, and is preferably 28 DEG C.
Wherein, the time of described restoration ecosystem is 4-7 days, is preferably 7 days.
Wherein, the described cotton cotyledon phase resists cold authentication method, carries out under the normal lighting conditions of cotton planting, and described normal lighting conditions is preferably illumination/dark for 14h/10h.
Wherein, described sub zero treatment, concrete operation method is: healthy and strong for the cotyledon period cotton planted in sand seedling is watered sufficient water, makes the water cut of sand reach mass percentage 85-95%, process under putting into low temperature environment.
Wherein, the healthy and strong seedling of the cotyledon period cotton of described kind in sand, for planting the healthy and strong seedling in the germination box taking sand as matrix, moisturizing added a cover by germination box.
The cotton cotyledon phase of the present invention resists cold authentication method, be preferably the healthy and strong seedling of the uniform strain cotyledon period to be measured of 30 strain growing way, 4 DEG C of sub zero treatment 24 hours, restoration ecosystem 7 days at 28 DEG C again, then investigate the state of stem and cotyledon to be hurt situation, damage to plants caused by sudden drop in temperature rank by the standard determination of table 1, with the formula 1, formula 2 calculates the anti-cold finger number of kind to be measured, differentiates anti-cold rank by the standard of table 2.
The present invention also provides the described cotton cotyledon phase to resist the application of cold authentication method in cotton breeding.
The present invention also provides the described cotton cotyledon phase to resist the application of cold authentication method in cotton planting.
The cotton cotyledon phase of the present invention resists cold authentication method cotyledon period can be damaged to plants caused by sudden drop in temperature by the area that cotyledon is hurt to be divided into 8 ranks (0 grade-7 grades, concrete division methods is in table 1), to divide detailed, workable.According to the Chilling injury index damaging to plants caused by sudden drop in temperature rank and investigate out each kind (being), be converted into anti-cold finger number again, according to anti-cold finger number, the cold resistance of cotton cotyledon phase is divided into 5 levels (see table 2), there is easy, directly perceived, reliable, that the cycle is short advantage, effectively can carry out the cold resistance screening of cotton in seedling stage germ plasm resource and kind.Germplasm resource for cotton have that scale is large, quantity is many and material source in the feature of the various variation types of planted in different ecological areas, so select suitable chilling tolerance detaction method particularly important, and cold resistance evaluation index really rule be weigh chilling tolerance detaction method whether fruitful key, the present invention just in time can meet the needs of this respect.
Accompanying drawing explanation
Fig. 1 is the curve map of 4 DEG C of sub zero treatment different times, 4 cotton material conductivity variations in preliminary experiment 2 of the present invention.
Fig. 2 is the reference photo that in the embodiment of the present invention 1, cotyledon period blade damages to plants caused by sudden drop in temperature rank criterion.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
Cotton material in all cotton variety the application involved in the application all comes from the Chinese Academy of Agriculture Science and Technologys Cotton Research Institute, be public kind, the public commercially or can introduce a fine variety from corresponding breeding units or the Chinese Academy of Agriculture Science and Technologys Cotton Research Institute and obtain.
The screening of preliminary experiment 1 sub zero treatment temperature
1, material: 4 upland cotton floral materials (title and source are in table 3).
Table 3 germination period resists kind (being) title and source needed for cold evaluation and screening
2, disposal route: 15 DEG C, 10 DEG C, 4 DEG C process 7 days respectively
3, investigate: the seedling rate alive after 7 days of recovering under 28 DEG C of conditions after sub zero treatment is in table 4.
The process of table 4 Different hypothermia recovers again the seedling rate/% alive after 7 days in 7 days
| Kind (being) title | 15℃ | 10℃ | 4℃ |
| Henan 2067 | 100.00 | 100.00 | 7.14 |
| sGK958 | 100.00 | 100.00 | 0.00 |
| GK50 | 100.00 | 100.00 | 15.38 |
| Cotton No. 3 of weighing apparatus | 100.00 | 100.00 | 10.71 |
4, result: as can be seen from 4,4 materials are under 15 DEG C, 10 DEG C sub zero treatment conditions, after restoration ecosystem, although growth is suppressed, the symptom that is hurt is not obvious, and seedling rate of living is 100%, restoration ecosystem 7 days after processing 7 days at 4 DEG C, 4 cotton materials are all subject to obvious injury, and seedling rate of living reduces, greatly so 4 DEG C of low temperature can as the first-selected condition of sub zero treatment.
The screening of preliminary experiment 2 sub zero treatment time
From preliminary experiment 1, under 4 DEG C of sub zero treatment conditions, upland cotton is subject to obvious injury, so 4 DEG C of sub zero treatment can as the first-selected condition of sub zero treatment, but process 7 days, 4 cotton material death rates reduce greatly, and the injury be subject to is all too strong, be unfavorable for the cold resistance distinguishing storeroom, so the concrete time of 4 DEG C of sub zero treatment also should be screened further.
1, material
In table 3.
2, disposal route: 4 DEG C of process 0h, 2h, 6h, 10h, 24h
3, investigation method
Cotyledon 0.5 gram is respectively got in each process of each material, shreds and adds ultrapure water 100ml, mixing, surveys conductivity (μ S/cm) after 28 DEG C of immersion 24h.
4, result investigation
4 DEG C of sub zero treatment different times, 4 cotton material conductivity variations are shown in Fig. 1.
The increase of conductivity illustrates the degree that is hurt of plant tissue cell's film, and conductivity is larger, illustrate be hurt heavier.As seen from Figure 1, except Henan 2067, all the other 3 materials are along with the prolongation of sub zero treatment time, conductivity is all in rising trend, what wherein GK50 and weighing apparatus cotton No. 3 rose is all very fast, and illustrating that the cell membrane of these two materials is hurt relatively more serious, is secondly sGK958, the conductivity variations in Henan 2067 is relatively more steady all the time, illustrates that its cotyledon cell film is smaller by low temperature effect.
During 4 DEG C of sub zero treatment 0h and 2h, the equal difference of conductivity of 4 cotton materials is not remarkable, during process 6h, the conductivity of 4 storerooms is pole significant difference, until difference is maximum during process 24h, as can be seen here, 4 DEG C of process 6h to 24h all can distinguish cold resistance between cotton material, and it is best to distinguish effect during process 24h.
The screening of preliminary experiment 3 sub zero treatment strain number
What process strain number represented is repeat number, and strain number is more, illustrates that repeat number is more, result is more accurate, but strain number is too many, and workload is too large again, so according to actual conditions, selected plant number 10 strain to 30 strain all can react the cold resistance of surveyed kind more exactly, and workload again can not be excessive.
The screening of preliminary experiment 4 restoration ecosystem temperature and time
It is 28 DEG C that cotton is applicable to growth temperature, so the temperature of restoration ecosystem elects 28 DEG C as.
4 DEG C of process 24h, 28 DEG C of restoration ecosystem injury symptom of 4 days to 7 days occurs all, so control time is restoration ecosystem 4 days to 7 days, but obvious not as 7 days of the symptom of 4 days, so in order to easily distinguish between different materials, the be hurt symptom of same material not between homophyletic, select restoration ecosystem to investigate for 7 days, be more conducive to efficiently identifying simultaneously.
Embodiment 1
1, the selection of material
Upland cotton cotyledon period resists the screening material therefor of cold identification of indicator to select 47 upland cotton flower varieties of current spread, these materials in field, seedling stage, sow in medium preliminary screening in advance, the Comprehensive Traits such as Agronomic characteristic are excellent, disease-resistant, pest-resistant, output can ensure carrying out smoothly of follow-up laboratory experiment.Its source and pedigree, in table 5, derive from the ground such as North China, Central China, Shandong of the U.S. and China, wide material sources, rich hereditary variation respectively.
Table 5 cotyledon period resists cold qualification test material source used and pedigree
2, test method
Chilling tolerance detaction is carried out to raise variety, method is by healthy and strong seedling 4 DEG C of sub zero treatment 24 hours of 30 strain strain to be measured cotyledon period, restoration ecosystem 7 days at 28 DEG C again, then investigate the state of stem and cotyledon to be hurt area, rank is damaged to plants caused by sudden drop in temperature by the standard determination of table 1, with the formula one, formula two calculates the anti-cold finger number of kind to be measured, differentiates anti-cold rank by the standard of table 2.Concrete details of operation is as follows:
1) the selecting of seed, selects great-hearted seed, full, uniformity, unabroken cotton seeds 50, Westerner or photon, and the present embodiment seed used is the seed that land for growing field crops in 2014 newly gathers in the crops, Westerner.Each kind establishes three repetitions.
2) cultivation of cotyledon period cotton seedling: it is mass percentage 23% that the sand mixed thoroughly by high-temperature sterilization adds water to moisture, evenly be laid in the rectangle germination box of long 19.5cm, wide 14cm, dark 5cm, the thickness of sand is about 2cm, above (50) that counted evenly are laid in for the seed trying material, add a cover the sand of same humidity and thickness again, cover the lid of long 19.3cm, wide 13.8cm, dark 7cm, add a cover moisturizing, be positioned in the illumination box of 28 DEG C of temperature, light application time is set to 14h, interlunation 10h.After 7d, not open and flat for cotyledon, weak seedling and lopsided seedling are pulled out, leaves uniform, the healthy and strong seedling of growing way 30 in every box for subsequent experimental.
3) cryogenic treatment process: the seedling reserved is watered sufficient water, makes the water cut of sandy soil reach mass percentage 85-95%, adds a cover moisturizing.Be put in that temperature is 4 DEG C, illumination/dark is process 24h in the growth case of 14h/10h, take out, be directly put in that temperature is 28 DEG C, illumination/dark is restoration ecosystem 7d in the growth case of 14h/10h.
4) chilling tolerance detaction standard:
First investigate the state of stem, be divided into stem rotten disconnected intact with stem, again for the individual plant investigation cotyledon dropping situations that stem is intact, investigate the area that is hurt of the cotyledon that do not come off, it is X that record speck area accounts for this sheet leaf area ratio, record total speck area account for total leaf area ratio be Y (when only having a slice cotyledon not come off, Y=X; X=0 is considered as) when cotyledon is intact; Fig. 2 is shown in by the reference photo of each rank concrete.
To be hurt situation according to the investigation state of stem and cotyledon, to damage to plants caused by sudden drop in temperature rank by the standard determination of table 1:
Table 1 upland cotton cotyledon period blade damages to plants caused by sudden drop in temperature rank
With the formula 1, formula 2 calculates the anti-cold finger number of kind to be measured:
Chilling injury index (%)=∑ (rank × this grade of strain number)/(8 × total strain number) × 1001
Anti-cold finger number (%)=100-Chilling injury index 2
According to the anti-cold finger number calculating gained, differentiate anti-cold rank by the standard of table 2:
Table 2 cotyledon period resists cold qualification rank and evaluation criterion
| Anti-cold finger number/% | Anti-cold rank |
| 0.00-29.90 | Cold sensitivity |
| 30.00-49.90 | Not anti-cold |
| 50.00-74.90 | Cold-resistant |
| 75.00-89.90 | Anti-cold |
| ≥90.00 | High resistance is cold |
。
3, test findings
After 4 DEG C of sub zero treatment 24h, between different materials and between the Different Individual of same material, damage to plants caused by sudden drop in temperature significant difference, show and all meet see Fig. 2 the sight that table 1 lists, and can obviously distinguish, illustrate and utilize these treatment conditions to identify that cotton cotyledon phase cold resistance is feasible.Test findings is in table 6:
The screening of table 6 cotton cotyledon phase cold resistance germ plasm resource
As can be seen from Table 6, cotyledon period cold resistance significant difference between different upland cotton seeds of flowering plants material, the reliability of this authentication method of sufficient proof.By 14.29% to 86.35% not etc., average anti-cold finger number is 51.60% for the anti-cold finger number of 47 parts of land Cotton Germplasms resource materials.In 47 parts of Germplasm Resources of Upland Cotton materials, cotyledon period resists cold material to have 8 parts, be respectively Handan 109,3268F6 is mixed in Henan 2067, GZ1, wound cotton 22, Ji, CCRI 44, middle 404A are anti-13, light seed 2, account for 17.02% of total expert evidence, cold-resistant has 16 parts, accounts for 34.04%, and anti-cold have 20 parts, cold sensitive material 3 parts, does not have the material that high resistance is cold.As can be seen here, identify that upland cotton cotyledon period cold resistance is feasible with 4 DEG C of process 24h.In long-term northern high latitude plantation screening training process, portion of material is had to have certain cold resistance (17.02%) in upland cotton, but for the cotton of cotyledon period, still have the material of half nearly still not anti-cold or cold sensitivity (42.55%).The differential screening of these materials be after the anti-cold breeding of cotton and the further research of anti-cold reason pave the way.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. a cotton cotyledon phase resists cold authentication method, it is characterized in that, it is by the healthy and strong seedling sub zero treatment of uniform for growing way many strains strain to be measured cotyledon period, restoration ecosystem at a normal temperature again, then investigate the state of stem and cotyledon to be hurt situation, damage to plants caused by sudden drop in temperature rank by the standard determination of table 1, with the formula 1, formula 2 calculates the anti-cold finger number of kind to be measured, differentiates anti-cold rank by the standard of table 2;
The state of described investigation stem and cotyledon are hurt area, specific as follows: the state first investigating stem, be divided into stem rotten disconnected intact with stem, again for the individual plant investigation cotyledon dropping situations that stem is intact, investigate the area that is hurt of the cotyledon that do not come off, it is X that record speck area accounts for this sheet leaf area ratio, and recording total speck area, to account for total leaf area ratio be Y;
Table 1 upland cotton cotyledon period blade damages to plants caused by sudden drop in temperature rank
Chilling injury index (%)=∑ (rank × this grade of strain number)/(8 × total strain number) × 1001
Anti-cold finger number (%)=100-Chilling injury index 2
Table 2 cotyledon period resists cold qualification rank and evaluation criterion
2. authentication method according to claim 1, is characterized in that, described many strains be 10 strains and more than.
3. authentication method according to claim 1, is characterized in that, the described sub zero treatment of process wherein, and treatment temperature is 0-15 DEG C, and the processing time is 6-24h.
4. authentication method according to claim 3, is characterized in that, the described sub zero treatment of process wherein, and treatment temperature is 4 DEG C, and the processing time is 24h.
5. authentication method according to claim 1, is characterized in that, the time of described restoration ecosystem is 4-7 days.
6. authentication method according to claim 1, it is characterized in that, described sub zero treatment, concrete operation method is: healthy and strong for the cotyledon period cotton planted in sand seedling is watered sufficient water, make the water cut of sand reach mass percentage 85-95%, process under putting into low temperature environment.
7. authentication method according to claim 6, is characterized in that, the healthy and strong seedling of the cotyledon period cotton of described kind in sand, for planting the healthy and strong seedling in the germination box taking sand as matrix, moisturizing added a cover by germination box.
8. the method according to any one of claim 1 ~ 7, it is characterized in that, it is by the healthy and strong seedling of the uniform strain cotyledon period to be measured of 30 strain growing way, 4 DEG C of sub zero treatment 24 hours, restoration ecosystem 7 days at 28 DEG C again, then investigates the state of stem and cotyledon and to be hurt situation, damage to plants caused by sudden drop in temperature rank by the standard determination of table 1, with the formula 1, formula 2 calculates the anti-cold finger number of kind to be measured, differentiates anti-cold rank by the standard of table 2.
9. the cotton cotyledon phase described in any one of claim 1 ~ 8 resists the application of cold authentication method in cotton breeding.
10. the cotton cotyledon phase described in any one of claim 1 ~ 8 resists the application of cold authentication method in cotton planting is promoted.
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| CN201510948563.8A CN105447763B (en) | 2015-12-17 | 2015-12-17 | A kind of cotton cotyledon phase resists cold identification method |
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| CN105447763B CN105447763B (en) | 2019-11-26 |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105875225A (en) * | 2016-04-18 | 2016-08-24 | 山西省农业科学院小麦研究所 | Method for identifying spring frosting resistance of wheat by utilizing leaf freezing injury |
| CN107182513A (en) * | 2017-05-16 | 2017-09-22 | 广东省农业科学院蔬菜研究所 | Wax gourd Evaluation of Cold Tolerance method |
| CN108575645A (en) * | 2018-04-03 | 2018-09-28 | 中国农业科学院棉花研究所 | A kind of method of field Fast Evaluation cotton to herbicide tolerant |
| CN109258371A (en) * | 2018-09-04 | 2019-01-25 | 上海市农业科学院 | A kind of evaluation method of florescence red palm cold resistance |
| CN110073974A (en) * | 2019-06-04 | 2019-08-02 | 中国农业科学院棉花研究所 | It is a kind of to screen the method and its application for resisting cold cotton variety |
| CN110235639A (en) * | 2019-05-20 | 2019-09-17 | 中国农业科学院棉花研究所 | Nature seawater coerces cotton in seedling stage Tolerant salt method |
| CN113748946A (en) * | 2021-09-14 | 2021-12-07 | 中国农业科学院棉花研究所 | Method for identifying cold resistance of upland cotton |
-
2015
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Non-Patent Citations (1)
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| 王俊娟等: "棉花萌发期抗冷性鉴定", 《中国农学会棉花分会会议论文集》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105875225A (en) * | 2016-04-18 | 2016-08-24 | 山西省农业科学院小麦研究所 | Method for identifying spring frosting resistance of wheat by utilizing leaf freezing injury |
| CN107182513A (en) * | 2017-05-16 | 2017-09-22 | 广东省农业科学院蔬菜研究所 | Wax gourd Evaluation of Cold Tolerance method |
| CN108575645A (en) * | 2018-04-03 | 2018-09-28 | 中国农业科学院棉花研究所 | A kind of method of field Fast Evaluation cotton to herbicide tolerant |
| CN109258371A (en) * | 2018-09-04 | 2019-01-25 | 上海市农业科学院 | A kind of evaluation method of florescence red palm cold resistance |
| CN110235639A (en) * | 2019-05-20 | 2019-09-17 | 中国农业科学院棉花研究所 | Nature seawater coerces cotton in seedling stage Tolerant salt method |
| CN110073974A (en) * | 2019-06-04 | 2019-08-02 | 中国农业科学院棉花研究所 | It is a kind of to screen the method and its application for resisting cold cotton variety |
| CN113748946A (en) * | 2021-09-14 | 2021-12-07 | 中国农业科学院棉花研究所 | Method for identifying cold resistance of upland cotton |
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| CN105447763B (en) | 2019-11-26 |
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